ZA200405136B - Intermediates for lhrh antagonist synthesis process for their production and process for lhrh antagonist production - Google Patents
Intermediates for lhrh antagonist synthesis process for their production and process for lhrh antagonist production Download PDFInfo
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- ZA200405136B ZA200405136B ZA2004/05136A ZA200405136A ZA200405136B ZA 200405136 B ZA200405136 B ZA 200405136B ZA 2004/05136 A ZA2004/05136 A ZA 2004/05136A ZA 200405136 A ZA200405136 A ZA 200405136A ZA 200405136 B ZA200405136 B ZA 200405136B
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- South Africa
- Prior art keywords
- boc
- 3pal
- lys
- 2nal
- heptapeptide
- Prior art date
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- 238000004519 manufacturing process Methods 0.000 title claims abstract description 12
- 238000000034 method Methods 0.000 title claims description 25
- 230000008569 process Effects 0.000 title claims description 23
- 229940124041 Luteinizing hormone releasing hormone (LHRH) antagonist Drugs 0.000 title claims description 21
- 239000002474 gonadorelin antagonist Substances 0.000 title claims description 15
- 230000015572 biosynthetic process Effects 0.000 title description 13
- 238000003786 synthesis reaction Methods 0.000 title description 11
- 239000000543 intermediate Substances 0.000 title description 9
- URKWHOVNPHQQTM-OAHLLOKOSA-N (2r)-2-[(2-methylpropan-2-yl)oxycarbonylamino]-3-naphthalen-2-ylpropanoic acid Chemical compound C1=CC=CC2=CC(C[C@@H](NC(=O)OC(C)(C)C)C(O)=O)=CC=C21 URKWHOVNPHQQTM-OAHLLOKOSA-N 0.000 claims abstract description 4
- 150000003839 salts Chemical class 0.000 claims abstract 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 21
- 150000001413 amino acids Chemical class 0.000 claims description 18
- 125000006239 protecting group Chemical group 0.000 claims description 15
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 10
- -1 Boc group Chemical group 0.000 claims description 7
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 claims description 6
- 150000001875 compounds Chemical class 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- XBPCUCUWBYBCDP-UHFFFAOYSA-N Dicyclohexylamine Chemical compound C1CCCCC1NC1CCCCC1 XBPCUCUWBYBCDP-UHFFFAOYSA-N 0.000 claims description 3
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 claims description 3
- 125000002252 acyl group Chemical group 0.000 claims description 3
- RPACBEVZENYWOL-XFULWGLBSA-M sodium;(2r)-2-[6-(4-chlorophenoxy)hexyl]oxirane-2-carboxylate Chemical compound [Na+].C=1C=C(Cl)C=CC=1OCCCCCC[C@]1(C(=O)[O-])CO1 RPACBEVZENYWOL-XFULWGLBSA-M 0.000 claims 1
- 239000000203 mixture Substances 0.000 description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 13
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 12
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 12
- XLXSAKCOAKORKW-UHFFFAOYSA-N gonadorelin Chemical compound C1CCC(C(=O)NCC(N)=O)N1C(=O)C(CCCN=C(N)N)NC(=O)C(CC(C)C)NC(=O)CNC(=O)C(NC(=O)C(CO)NC(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C(CC=1NC=NC=1)NC(=O)C1NC(=O)CC1)CC1=CC=C(O)C=C1 XLXSAKCOAKORKW-UHFFFAOYSA-N 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 238000010511 deprotection reaction Methods 0.000 description 6
- 238000000605 extraction Methods 0.000 description 5
- 238000001556 precipitation Methods 0.000 description 5
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 description 4
- VIHYIVKEECZGOU-UHFFFAOYSA-N N-acetylimidazole Chemical compound CC(=O)N1C=CN=C1 VIHYIVKEECZGOU-UHFFFAOYSA-N 0.000 description 4
- 230000008878 coupling Effects 0.000 description 4
- 238000010168 coupling process Methods 0.000 description 4
- 238000005859 coupling reaction Methods 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 229960004592 isopropanol Drugs 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 102000012673 Follicle Stimulating Hormone Human genes 0.000 description 3
- 108010079345 Follicle Stimulating Hormone Proteins 0.000 description 3
- 102000009151 Luteinizing Hormone Human genes 0.000 description 3
- 108010073521 Luteinizing Hormone Proteins 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000021736 acetylation Effects 0.000 description 3
- 238000006640 acetylation reaction Methods 0.000 description 3
- 229940028334 follicle stimulating hormone Drugs 0.000 description 3
- 229940040129 luteinizing hormone Drugs 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 238000010791 quenching Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- BTANRVKWQNVYAZ-SCSAIBSYSA-N (2R)-butan-2-ol Chemical compound CC[C@@H](C)O BTANRVKWQNVYAZ-SCSAIBSYSA-N 0.000 description 2
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 description 2
- 101000904173 Homo sapiens Progonadoliberin-1 Proteins 0.000 description 2
- 102100024028 Progonadoliberin-1 Human genes 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 101000996723 Sus scrofa Gonadotropin-releasing hormone receptor Proteins 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 2
- 125000004185 ester group Chemical group 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 239000012074 organic phase Substances 0.000 description 2
- 239000011541 reaction mixture Substances 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 206010004446 Benign prostatic hyperplasia Diseases 0.000 description 1
- 101100086315 Chlamydomonas reinhardtii RAA3 gene Proteins 0.000 description 1
- 208000005171 Dysmenorrhea Diseases 0.000 description 1
- 206010013935 Dysmenorrhoea Diseases 0.000 description 1
- 201000009273 Endometriosis Diseases 0.000 description 1
- 108700012941 GNRH1 Proteins 0.000 description 1
- 239000000579 Gonadotropin-Releasing Hormone Substances 0.000 description 1
- OTXBNHIUIHNGAO-UWVGGRQHSA-N Leu-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(O)=O)CCCCN OTXBNHIUIHNGAO-UWVGGRQHSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 238000009903 catalytic hydrogenation reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000010931 ester hydrolysis Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 108010034529 leucyl-lysine Proteins 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- 230000006340 racemization Effects 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/08—Tripeptides
- C07K5/0802—Tripeptides with the first amino acid being neutral
- C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/23—Luteinising hormone-releasing hormone [LHRH]; Related peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/02—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length in solution
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/08—Tripeptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/08—Tripeptides
- C07K5/0827—Tripeptides containing heteroatoms different from O, S, or N
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Endocrinology (AREA)
- Analytical Chemistry (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Nitrogen Condensed Heterocyclic Rings (AREA)
Abstract
The invention relates to a process for preparing a tripeptide, including a salt thereof, of the formula (IX)
Boc-D-2Nal-D-4CIPhe-D-3Pal-OH (IX)
comprising the following consecutive steps: (a) Reacting Boc-D-4ClPhe-OH with HONSu to form Boc-D-4ClPhe-OSu (VII); (b) Reacting Boc-D-4ClPhe-OSu (VII) with H-D-3Pal-OH to form Boc-D-4ClPhe-D-3Pal-OH (VIII); (c) Reacting Boc-D-4ClPhe-D-3Pal-OH (VIII) with Boc-D-2Nal-OSu prepared by reacting Boc-D-2Nal-OH with HONSu to form Boc-D-2Nal-D-4ClPhe-D-3Pal-OH (IX).
Description
. INTERMEDIATES FOR LHRH ANTAGONIST SYNTHESIS, PROCESS FOR - THEIR PRODUCTION, AND PROCESS FOR LHRH ANTAGONIST PRODUCTION
The present invention relates to intermediates for the synthesis of LHRH antagonists, to a process for the production of these intermediates and to a process for the production of LHRH antagonists.
The luteinizing hormone-releasing hormone, LHRH, controls the secretion of follicle stimulating hormone (FSH) and luteinizing hormone (LH). LHRH antagonists are compounds capable of blocking the secretion of FSH and LH. They are generally nona- and decapeptides (but may be shorter or longer) comprising part of or the entire structure of LHRH in which one or several amino acids have been exchanged for other natural amino acids and/or amino acids not found in nature.
Synthetic LHRH antagonists may be used for contraception and in the treatment of benign hyperplasia of the prostate gland, hormonal-dependent tumors of the breast and ovaries, dysmenorrhea, endometriosis, and other conditions. These synthetic LHRH antagonists have the general formula
Ac~D-2Nal-D-4C1lPhe-D-3Pal-Ser—-X-NHj,
® WO 03/055902 PCT/TB02/05583 wherein X is from 5 to 6 natural and/or synthetic amino acid ) residues. More particularly they have the aforementioned general formula wherein X is AAl-AA2-Leu-AA3-Pro-D-Ala, in particular wherein AAl is a natural or synthetic amino acid and AA2 is a natural or synthetic amino acid or zero, RAA3 is a natural or synthetic amino acid.
While there are a number of synthetic methods for preparing
LHRH analogs known in the art, there is a need for improvement since the total yield of LHRH analogs obtained from known processes is not high and the products, in addition, may require extensive purification. Moreover, the methods for the synthesis of LHRH analogs known in the art are quite costly.
A synthesis strategy disclosed in U.S. patent no. 5,710,246 for making decapeptide or nonapeptide LHRH antagonists ) comprises the coupling of an intermediate tripeptide representing amino residues 1 to 3 (counting starts at the amino terminal of the peptide) with a heptapeptide or a hexapeptide, respectively representing amino acid residues 4- 10 and 4-9, respectively. The intermediate tripeptide disclosed in US 5710246 A is an ester, Boc-D-2Nal-D-4ClPhe-D- 3Pal-0-Me or the corresponding benzyl or allyl ester.
It is thus an object of the invention to provide a tripeptide intermediate for the 3+7 and 3+6 synthesis of LHRH analogs in which the yield and/or purity of the product is improved.
It is another object of the invention to provide a process for the production of such a tripeptide intermediate.
® WO 03/055902 PCT/IB02/05583 : It is still another object of the-invention to provide a ’ process for the production of LHRH analogs in which a tripeptide is coupled to a hepta- or hexapeptide.
Further objects of the invention will become obvious from the following summary of the invention, the description of preferred embodiments, and the appended patent claims.
DEFINITIONS AND ABBREVIATIONS
For definitions and abbreviations used in this application and which are generally accepted in the field of the invention reference is made in particular to Us 5710246 A.
According to the invention is provided a tripeptide representing amino acids 1-3 of an LHRH antagonist, the terminal amino group of which is Boc- or Ac-protected and the terminal carboxyl group of which (that is, the terminal group of amino acid no. 3) is not protected.
According to the invention is disclosed the tripeptide (I)
Ac-D-2Nal-D-4C1Phe-D-3Pal-OH (I) which is a useful intermediate in a process for the synthesis of an LHRH antagonist of the general formula (II)
Ac-D-2Nal-D-4ClPhe-D-3Pal-Ser-X-NH; (II)
® WO 03/055902 PCT/IB02/05583 wherein X is from 5 to 7 natural and/or synthetic amino acid ’ residues, more preferred AAl-AA2-Leu-AA3-Pro-D-Ala, in particular wherein AAl is a natural or synthetic amino acid and BA? is a natural or synthetic amino acid or zero, AA3 is a natural or synthetic amino acid.
Still preferred is the use of the tripeptide (I) in the synthesis of a peptide of the general formula (IIa)
Ac-D-2Nal-D-4ClPhe-D-3Pal-Ser-AAl-AA2-Leu-Lys (iPr)~Pro-D-Ala-
NH; (IIa), wherein AA1l and AA2 have the meaning given above, in particular a LHRH antagonist of the formula (III)
Ac-D-2Nal-D-4ClPhe-D-3Pal-Ser-MeTyr-D-Lys (Nic) ~Leu-Lys (iPr) -
Pro-D-Ala-NH, (III) or, even more preferred, of the formula (IIIa)
Ac-D-2Nal-D-4ClPhe-D-3Pal-Ser-MeTyr-D-Asn-Leu-Lys (iPr) -Pro-D-
Ala-NH; (IIIa).
According to the invention is also disclosed the tripeptide
Boc-D-2Nal-D-4ClPhe-D-3Pal-OH (IX) : of same utility.
Furthermore, according to the invention is disclosed a process for preparing a tripeptide of the formula (I)
Ac-D-2Nal-D-4ClPhe~-D~3Pal-OH (I)
® WO 03/055902 PCT/IB02/05583 or (IX) . .
Boc-D-2Nal-D-4ClPhe-D-3Pal-OH (IX), 5 comprising the following consecutive steps for the preparation of (I): (a) Reacting Boc-D-4ClPhe-OH with HONSu to form
Boc—-D-4C1lPhe-0Su (VII); (b) Reacting Boc-D-4ClPhe-OSu (VII) with H-D-3Pal-OH to form
Boc-D-4ClPhe-D-3Pal-0H (VIII); (c) Reacting Boc-D-4ClPhe-D-3Pal-OH (VIII) with Boc-D-2Nal-
OSu prepared by reacting Boc-D-2Nal-OH with HONSu to form
Boc-D-2Nal-D-4ClPhe~-D-3Pal-OH (IX); (d) Reacting Boc-D-2Nal-D-4ClPhe-D-3Pal-OH (IX) with acetic acid to form Ac-D-2Nal-4ClPhe-D-3Pal-OH (I); or the consecutive steps (a) through (c) for the preparation of (IX).
The process of the invention for preparing a LHRH antagonist comprises the step of coupling the tripeptide (I) with a heptapeptide (IV) of the general formula
Pl-Ser (P?) -AAl-AA2-Leu-Lys (iPr, P*) -Pro-D-AlaNH, (IV), wherein P* is H or an amino protecting group such as Boc, wherein AAl and 2A2 have the aforementioned meaning, in particular with a heptapeptide (V) of the general formula pl-Ser (P?) ~NMeTyr (P?) -D-Lys (Nic) -Leu-Lys (iPr, P*) -Pro~D—-AlaNH, (V), wherein P! is selected from H or amino protecting group and P? and P? are independently selected from H and -OH protecting group, and P?* has the meaning given above, for preparing the LHRH antagonist Ac-D-2Nal-D-4ClPhe-D-3Pal-Ser-—
MeTyr-D-Lys (Nic) -Leu~Lys (iPr) -Pro-D-Ala-NH; (111),
® WO 03/055902 PCT/IB02/05583 more particularly with a heptapeptide (Va) of the general formula P'-Ser (P?)-NMeTyr (P®)~D-Asn-Leu-Lys (iPr, P*) -Pro-D—
AlaNH, (Va), wherein P! is selected from H or amino protecting group and P? and P? are independently selected from H and —OH protecting group, and P' has the meaning given above, for preparing the LHRH antagonist Ac-D-2Nal-D-4ClPhe-D-3Pal~Ser-
MeTyr-D-Asn-Leu- Lys (iPr)-Pro-D-Ala-NH (III).
The heptapeptide (V) is described in US 5710246 A. The heptapeptide of the general formula (IV) including the heptapeptide (Va) can be synthesized by routine modifications of the synthesis of (V) or by coupling the corresponding Boc- amino acids on a peptide synthesizer (Beckman Model 990), as described in WO 94/40757 where also the LHRH antagonist (III) is disclosed.
Alternatively the process of the invention for preparing a : LHRH antagonist comprises the step of coupling the tripeptide . (IX)
Co © Boc-D-2Nal-D-4ClPhe-D-3Pal-OH (IX) with a heptapeptide (IV) of the general formula pl-Ser (P?)-AAl-AA2-Leu-Lys (iPr, P?) -Pro-D-AlaNH: (IV), wherein P!, P2, PY AAl and AA2 have the meaning given above, in particular with a heptapeptide (V) of the general formula p!-Ser (P?) -NMeTyr (P?) -D-Lys (Nic) -Leu-Lys (iPr, P!) -Pro-D—AlaNH, (Vv) or, even more preferred, with a heptapeptide of the general formula (Va)
® WO 03/055902 PCT/IB02/05583
P'-Ser (P?) -NMeTyr (P®) -D-Asn-Leu-Lys (iPr, P*) -Pro-D-AlaNH; (Va), wherein p! is selected from BE or amino protecting group, p? and P° are independently selected from H and —OH protecting group, Pp? has the aforementioned meaning, followed by substituting the N-terminal Boc group by an acyl group, in particular an acetyl group. )
More particularly, the heptapeptide of the general formula (V) is the heptapeptide (VI)
H-Ser (tBu) -NMeTyr-D-Lys (Nic) -Leu-Lys (iPr, Boc) ~Pro-D—-AlaNH; ¢ (Vi), or even more preferred, the heptapeptide (VIa)
H-Ser (tBu) -NMeTyr-D-Asn-Leu-Lys (iPr, Boc) -Pro-D—AlaNH; (VIa).
A particular advantage with the method of the invention is that a cheaper starting material, E-D-Pal-OH-2HC1l, can be oo used instead of the ester H-Pal-OR-2HCl; the protective group of the starting material need not be removed. Therefore the synthesis of the invention is one step shorter and avoids rhat material is lost in the additional step. Another advantage is that the formation of impurities in the saponification step is avoided. The formation of such impurities is well known. For instance, the basic conditions at the ester hydrolysis step cause partial racemization of
D-Pal. The other prior-art alternative of removing the ester group by catalytic hydrogenation (in the case of allyl or benzyl ester groups) risks to cause a loss of Cl from 4ClPhe producing Phe. While allyl groups may be removed by still other reagents the full removal is difficult to control.
® WO 03/055902 PCT/IB02/05583
The invention will now be explained in more detail by : reference to a preferred embodiment.
] Synthesis of Ac-D-2Nal-4ClPhe-D-3Pal-OH (I). .
EXAMPLE 1. Boc-D-4ClPhe-0Su. Boc-D-4ClPhe-OH (299,75 g; 1,0 eq.) and ‘HONSu (184,1 g; 1,6 eq.) are dissolved in 2-propanol (4,5 1). The mixture is cooled to 0°C and DIC {164,1 g; 1.3 eq.) is added. The mixture is stirred for 16h while warming to room temperature. The product is filtered of, washed with 2-propanocl (1,5 L) and dried. Yield: 85%. HPLC purity: 98,83.
EXAMPLE 2. Boc-D-4ClPhe-D-3Pal-OH. H-D-3Pal-OH, 2 HCl (251,1 g; 1,05 eq.) and Boc-D-4ClPhe-OSu (396,8 g; 1,0 eg.) are dissolved in DMSO (3,33 L) and NMM (318,8 g; 3,15 eq:) is added. The mixture is stirred for 16 h at room temperature.
Water (17 L) is added and pH is adjusted to 4-4,5 which causes the product to precipitate. The mixture is filtered and the product is washed with water (3 Xx 5 L) to remove traces of DMSO, H-D-3Pal-OH and Boc-D-4ClPhe-OH. The product is dried. Yield: 80%. HPLC purity: 97,8% :
EXAMPLE 3. Boc-D-2Nal-OSu. Boc-D-2Nal-OH (315,4 g¢g; 1,0 eq.) is dissolved in 2-propanol (6,8 L) ‘at ~10°C and IBC (157 g; 1,15 eq.) and NMM (116 g; 1,15 eq.) is added. After stirring for 5-10 min a mixture of HONSu (230,11 g; 2,0 eq.) in 2- propanol (1,4 L) is added. Additional NMM (10,1 g; 0,1 eg.) is added. After half an hour water (0,82 L) is added to dissolve precipitated NMM-HCl. The product is isolated by
® WO 03/055902 PCT/IB02/05583
X 9 filtration, washed with 2-propanol (1 L), and dried. Yield: ’ 90%. HPLC purity: 98, 3%.
EXAMPLE 4. Boc-D-Nal-D-4ClPhe-D-3Pal-OH. (a) Deprotection. Boc-D-4ClPhe-D-3Pal-OH (447,93 g; 1,0 eq.) is dissolved in a mixture of ethyl acetate (3,4 L), acetic acid (675 ml) and MSA (454 mL; 7,0 eq.) at 0°C and kept at this temperature for two hours. TEA (1669 ml; 12 eq.) is added. b) Condensation. Boc-D-Nal-OSu (412,4 g; 1,0 eq.) 1s added to the neutralized deprotection mixture at room temperature. The reaction mixture is kept at this temperature for 2-4 h. Aq. 25% NH; (154 mL; 2,0 eq.) is added to quench remaining hydroxysuccinimide ester. 1-Butanol (4,5 L) is added to prevent precipitation in the subsequent extractions. ¢) Purification and isolation. The reaction mixture is extracted twice at pH 6 (2 x 4,5 L water) to remove TEA, at pH 9 (4,5 L water) to remove MSA and finally at pH 7 (4,5 L water). The extractions are carried out at 40-45°C to prevent precipitation. To the organic phase is added acetic acid (4,5 oo L; 1 vol.) and the mixture is concentrated in vacuo and co- evaporated with acetic acid (4,5 L) to give a solid.
EXAMPLE 5. Ac-D-2Nal-D-4ClPhe-D-3Pal-ONa. . a) Deprotection. To the solid Boc-D-2Nal-D-4ClPhe-D-3Pal-OH is © added water (90 ml), acetic acid (1,8 L) and MSA (454 mL; 7,0 eq.) and the mixture is stirred for 1-2 h at room temperature.
The mixture is cooled to 0°C and neutralized with TEA (1071 ml; 7,7 eq.). The solution is concentrated in vacuo and co- evaporated twice with toluene (2 x 2,5 L) to give an oil. b) Acetylation. The oil from the deprotection step is dissolve in toluene (2,0 L) and acetyl imidazole (132,14 g) is added.
® WO 03/055902 PCT/1B02/05583
The mixture is stirred at room temperature for 1 h and then : water (100 ml) is added to quench remaining acetyl imidazole. d) Purification. The mixture from the acetylation is heated to 30-35°C and l-butanol (4,5 L) is added to prevent precipitation
The mixture is extracted twice at pH 5 (2 x 2,6 L water), and twice at pH 11 (2 x 2,6 L water) using NaOH to adjust pH to 11.
Methanol (2,25 L) is added to the last extractions to prevent precipitation. NaCl (130 g) is added to the first and the last extraction to minimize loss of product in the aqueous phases. e) Isolation. To the vigorously stirred organic phase from the extractions is added heptane (15 L) and the resulting suspension is left at room temperature while stirring for at least 1 h. The mixture is filtered and the product is washed twice with heptane (2 x 3,5 L) and dried. Yield: 75% (from Boc-
D-4ClPhe-D-3Pal-OH). HPLC purity: 92%. Amino acid analysis: 2Nal: 1.1; 4ClPhe: 1.0; 3Pal: 0.9. MS: MW 586. Na: 4.6%
EXAMPLE 6. Ac-D-2Nal-D-4C1Phe-D-3Pal-OH * DCHA a) Deprotection. To the solid Boc-D-2Nal-D-4ClPhe-D-3Pal-OH is added water (90 mL), acetic acid (1,8 L) and MSA (454 mL; 7,0 eq.) and the mixture is stirred for 1-2 h at room temperature. The mixture is cooled to 0°C and neutralized with
TEA (1071 mL; 7,7 eq.). The solution is concentrated in vacuo and co-evaporated twice with toluene (2 Xx 2,5 L) to give an oil. b) Acetylation. The oil from the deprotection is dissolved in toluene (2,0 L) and acetyl imidazole (132,14 g) is added. The mixture is stirred at room temperature for 1 h followed by addition of water (100 ml) to quench remaining acetyl imidazole. c) Purification. The mixture is heated to 30-35°C and l-butanol (4,5 L) is added to prevent precipitation. The mixture is extracted twice at pH 7 (2 x 2,6 L water), once at pH 9-9,5
Claims (19)
1. A process for preparing a tripeptide, including a salt thereof, of the formula (1) Ac-D-2Nal-D-4CIPhe-D-3Pal-OH (1) or Boc-D-2Nal-D-4ClPhe-D-3Pal-OH (IX) comprising the following consecutive steps for the preparation of (I): (a) Reacting Boc-D-4CIPhe-OH with HONSu to form Boc-D-4CiPhe- OSu (VII); (b) Reacting Boc-D-4CIPhe-OSu (VII) with H-D-3Pal-OH to form Boc- D-4CIPhe-D-3Pal-OH (Vill); (c) Reacting Boc-D-4CIPhe-D-3Pal-OH (VIII) with Boc-D-D2Nal-OSu prepared by reacting Boc-D-2Nal-OH “with HONSu to form Boc-D-2Nal-D- 4C|Phe-D-3Pal-OH (IX); (d) Reacting Boc-D-2Nal-D-4ClPhe-D-3Pal-OH (1X) with acetic acid to form Ac-D-2Nal-4ClPhe-D-3Pal-OH (1); : AMENDED SHEET
} 14 PCT/IB02/05583 - or the consecutive steps (a) through (c) for the preparation of (1X).
2. The process of claim 1 for further preparing a LHRH antagonist or a pharmaceutically acceptable salt thereof, wherein ] the tripeptide Ac-D-2Nal-D-4CiPhe-D-3Pal-OH (1) is coupled with a heptapeptide (IV) of the general formula P'-Ser (P?)-AA1-AA2-Leu-Lys (iPr, P*)-Pro-D-AlaNH; (IV), wherein P' is selected from H or amino protecting group, P2 is H or OH- protecting group, P* is H or an amino protecting group such as Boc, AA1 is natural or synthetic amino acid and AA2 is natural or synthetic amino acid or zero.
3. The process of claim 2, wherein the heptapeptide of the general formula (IV) is P'-Ser (P?)-NMeTyr (P?)-D-Lys(Nic)-Leu-Lys(iPr, P*)-Pro-D-AlaNH; (V) wherein P3 is H or —OH protecting group.
4, The process of claim 2, wherein the heptapeptide of the general formula (IV) is P'-Ser (P2)-NMeTyr (P%)-D-Asn-Leu-Lys(iPr, P*)-Pro-D-AlaNH; (Va) wherein P® is H or —OH protecting group.
5. The process of claim 3, wherein the heptapeptide of the general formula (V) is H-Ser(tBu)-NMeTyr-D-Lys(Nic)-Leu-Lys(iPr, Boc)-Pro-D-AlaNH; (VI). AMENDED SHEET
PCT/IB02/05583
=
6. The process of claim 4, wherein the heptapeptide of the formula (V1) is a heptapeptide of the formula H-Ser(tBu)-NMeTyr-D-Asn-Leu-Lys(iPr, Boc)-Pro-D-AlaNHz (Via).
7. The process of claim 1 for further preparing an LHRH antagonist or 5s a pharmaceutically acceptable salt thereof, wherein the tripeptide Boc-D-2Nal-D-4CIPhe-D-3Pal-OH (IX) is coupled with a heptapeptide (IV) of the general formula P'-Ser (P)-AA1-AA2-Leu-Lys (iPr, P*)-Pro-D-AlaNHz (IV), wherein P' is selected from H or amino protecting group, P2 is H or OH- protecting group,. P* is H or an amino protecting group such as Boc, AA1 is natural or synthetic amino acid and AA2 is natural or synthetic amino acid or 2ero.
8. The process of claim 7, wherein the heptapeptide of the general formula (IV) is P'-Ser (P?)-NMeTyr (P?)-D-Lys(Nic)-Leu-Lys (iPr, P*)-Pro-D-AlaNH; (V) wherein P? is H or —OH protecting group.
9. The process of claim 8, wherein the heptapeptide of the general formula (V) is H-Ser(tBu)-NMeTyr-D-Lys(Nic)-Leu-Lys (ipR, Boc)-Pro-D-AlaNH: (VI).
10. The process of claim 7, wherein the heptapeptide of the general formula (IV) is P'-Ser (P?)-NMeTyr (P?)-D-Asn-Leu-Lys (iPr, P*)-Pro-D-AlaNH; (Va), AMENDED SHEET
PCT/IB02/05583 followed by substituting the Boc group by an acyl group, in particular an acetyl group.
11. The process of claim 10, wherein the heptapeptide of the general formula (IV) is H-Ser(tBu)-NMeTyr-D-Asn-Leu-Lys(ipR,Boc)-Pro-D-AlaNH ,(Vla). followed by substituting the N-terminal Boc group by an acyl group, in particular an acetyl group.
12. A salt of the tripeptide Ac-D-2Nal-D-4ClPhe-D-3Pal-OH(l).
13. The tripeptide Boc-D-2Nal-D-4CIPhe-D-3Pal-OH (IX) or a salt thereof.
14. Use of the Na salt or the DCHA complex of the tripeptide Ac- D-2Nal-D-4CIPhe-D-3Pal-OH (I) for preparing a LHRH antagonist or a pharmaceutically acceptable salt thereof.
15. A process according to any one of claims 1 to 11, substantially as herein described and illustrated.
16. A salt according to claim 12, substantially as herein described and illustrated.
17. A compound according to claim 13, substantially as herein described and illustrated.
18. Use according to claim 14, substantially as herein described and illustrated. AMENDED SHEET
PCT/IB02/05583
19. A new process for preparing a compound, a new salt, a new compound, or a new use of a salt or DCHA complex as defined in claim 14, substantially as herein described. AMENDED SHEET
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SE0104463A SE0104463D0 (en) | 2001-12-29 | 2001-12-29 | Intermediates for Synthesis of LHRH Antagonists, Methods of Preparation and Methods for Preparation of LHRH Antagonists |
| PCT/IB2002/005583 WO2003055902A1 (en) | 2001-12-29 | 2002-12-23 | Intermediates for lhrh antagonist synthesis, process for their production, and process for lhrh antagonist production |
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| ZA200405136B true ZA200405136B (en) | 2005-07-27 |
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| ZA2004/05136A ZA200405136B (en) | 2001-12-29 | 2004-06-28 | Intermediates for lhrh antagonist synthesis process for their production and process for lhrh antagonist production |
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| Country | Link |
|---|---|
| US (2) | US20050124788A1 (en) |
| EP (2) | EP1630169B1 (en) |
| JP (1) | JP4272529B2 (en) |
| KR (1) | KR100844520B1 (en) |
| CN (1) | CN1304417C (en) |
| AT (2) | ATE342913T1 (en) |
| AU (1) | AU2002348749B2 (en) |
| CA (1) | CA2471723C (en) |
| CY (2) | CY1107580T1 (en) |
| DE (2) | DE60215544T2 (en) |
| DK (2) | DK1630169T3 (en) |
| ES (2) | ES2296050T3 (en) |
| HK (1) | HK1075052A1 (en) |
| IL (2) | IL162788A0 (en) |
| NO (1) | NO330129B1 (en) |
| NZ (1) | NZ534245A (en) |
| PT (2) | PT1465917E (en) |
| RU (1) | RU2307125C2 (en) |
| SE (1) | SE0104463D0 (en) |
| SI (1) | SI1630169T1 (en) |
| WO (1) | WO2003055902A1 (en) |
| ZA (1) | ZA200405136B (en) |
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| JP4407322B2 (en) | 2004-03-09 | 2010-02-03 | 味の素株式会社 | Method for producing peptide |
| CN101037472B (en) * | 2006-03-14 | 2013-03-27 | 中国人民解放军军事医学科学院毒物药物研究所 | LHRH antagonist with low-histamine releasing function |
| TWI539959B (en) | 2008-02-11 | 2016-07-01 | 菲瑞茵國際中心股份有限公司 | Method of treating metastatic stage prostate cancer |
| SI2632934T1 (en) | 2010-10-27 | 2017-02-28 | Ferring B.V. | Process for the manufacture of degarelix and its intermediates |
| EP2447276A1 (en) * | 2010-10-27 | 2012-05-02 | Ferring B.V. | Process for the manufacture of Degarelix and its intermediates |
| MY182320A (en) | 2012-06-01 | 2021-01-19 | Ferring Bv | Downstream process |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4098777A (en) * | 1977-03-14 | 1978-07-04 | Merck & Co., Inc. | Process for the preparation of pyroglutamyl-Ala-Lys-Ser-Gln-Gly-Gly-Ser-Asn |
| US5225598A (en) * | 1990-03-30 | 1993-07-06 | Union Carbide Chemicals & Plastics Technology Corporation | Selective production of linear pentaethylenehexamine and hydroxyethyldiethylenetriamine |
| EP0540626A1 (en) * | 1990-07-24 | 1993-05-12 | Eastman Chemical Company | Process for synthesizing peptides |
| US5710246A (en) * | 1996-03-19 | 1998-01-20 | Abbott Laboratories | Process for intermediates for the synthesis of LHRH antagonists |
| ES2264169T3 (en) * | 1996-10-30 | 2006-12-16 | Pfizer Inc. | AZABICICLIC DERIVATIVES OF PIRIDONA FUSIONADA OR CITISINA, ITS PREPARATION AND ITS USE IN THE ADDICTION THERAPY. |
| US5968895A (en) * | 1996-12-11 | 1999-10-19 | Praecis Pharmaceuticals, Inc. | Pharmaceutical formulations for sustained drug delivery |
| DE19941248A1 (en) | 1998-09-01 | 2000-03-02 | Asta Medica Ag | New cetrorelix analogs containing N-substituted glycine derivatives useful as LHRH antagonists, e.g. for treating hormone-dependent tumors |
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